Selection of murine lymphoid and hematopoietic cells using polystyrene tissue culture devices containing covalently immobilized antibody

Biotechniques. 1996 Jun;20(6):1098-103. doi: 10.2144/96206pf02.

Abstract

We have established rapid procedures that negatively deplete and positively select for specific murine cell populations. By using polystyrene tissue culture flasks containing a covalently bound mouse anti-rat antibody and specific anti-mouse, cell-surface antigen antibodies, we easily and efficiently depleted greater than 90% of the mature lineage cells from murine bone marrow. This selection procedure resulted in an enrichment of progenitor colonies (CFU-Cs) in murine bone marrow. Using the same polystyrene tissue culture devices, we can directly isolate CD117+ (c-kit+) murine hematopoietic cells. As few as 2000 of these CD117+ cells rescued and reconstituted lethally irradiated recipients in a murine bone marrow transplant model.

MeSH terms

  • Animals
  • Bone Marrow Transplantation
  • Cell Lineage
  • Cell Separation / methods*
  • Culture Techniques / instrumentation
  • Culture Techniques / methods*
  • Flow Cytometry / methods
  • Hematopoietic Stem Cells*
  • Lymphocytes / immunology
  • Mice
  • Mice, Inbred C57BL
  • Proto-Oncogene Proteins c-kit / immunology

Substances

  • Proto-Oncogene Proteins c-kit